The second-generation sequencing technology, also known as Next-generation sequencing technology, is named corresponding to the first-generation sequencing technology which is represented by Sanger sequencing method. The second-generation sequencing technology is represented by Roche/454 Pyrosequencing, Illumina/Solexa polymerase synthesis sequencing and ABI/SOLiD ligase sequencing, and their common characteristics are high sequencing throughput. Compared with these mainstream sequencing platforms, Complete Genomics (CG) sequencing platform with the highest throughput may produce 9.9 TB of data in each run, and its output may reach 50 Gb per hour, which is 10-25 times that of the mainstream of the sequencing platform. With respect to read length for haploidy, among the mainstream sequencing platforms, only the Illumina sequencer may achieve a read length of 8-10 kb to haploidy, while the CG sequencer may reach a read length greater than 99 kb. In addition, the CG sequencer may achieve accuracy up to 99.999%, better than other commercial sequencers. Thus, compared with the mainstream sequencing platforms, CG sequencing platform has its unique advantages.
In the process of constructing a nucleic acid sequencing library, it is generally necessary to introduce an adaptor with a known sequence for sequencing. However, it has been reported that the adaptor is ligated for library construction in such an existing way that not only ligating efficiency is not high enough, but also many by-products at low come along. In addition, as CG sequencing platform adopts a cyclic single-stranded library for sequencing, thus linear double-stranded libraries constructed by the mainstream sequencing platforms are not suitable for the CG sequencers. However, as to the method for constructing the cyclic single-stranded library for the nucleic acid sequencing, there is no literature has been reported so far.
Based on above situations, an adaptor with high ligating efficiency and accuracy is urgently required to be developed in the related art.